mRNA from frog corneal epithelium increases water permeability in Xenopus oocytes. 1995

T Horwich, and C Ibarra, and P Ford, and A Zamudio, and M Parisi, and O A Candia
Department of Ophthalmology, Mount Sinai School of Medicine, New York, NY 10029-6574, USA.

OBJECTIVE To investigate the existence of a water channel in the frog corneal epithelium by studying the osmotic water permeability (Pf) of Xenopus oocytes expressing the mRNA message from frog corneal epithelium. METHODS Total RNA was obtained from corneal epithelium by a single-step phase separation method, and poly A+ RNA was isolated using oligo-dT columns. This mRNA was injected into the oocytes. After a 48-hour incubation, oocyte volume changes elicited by a hypoosmotic solution were measured with a computerized video system. RESULTS Oocytes injected with 50 nl mRNA (1 microgram/microliter) showed a significant increase in Pf compared to water-injected controls (8.4 +/- 1.5 to 17.5 +/- 1.9 cm.sec-1 x 10(-4), P < 0.005). mRNA-injected oocytes exposed to a higher external [Cl-] showed a heightened permeability. Furthermore, Pf of oocytes exposed to a solution containing the recognized water-channel blocker HgCl2 was significantly lower than the Pf of mRNA-injected oocytes not exposed to HgCl2. CONCLUSIONS Evidence was found for a water channel in the frog corneal epithelium because oocytes injected with the epithelial mRNA manifested increased water permeability. The increase in water permeability was larger in the presence of external Cl- and was inhibited by HgCl2. This finding correlates with measurements of Pf in the intact epithelium in which apical Cl- induced an increase in transepithelial water permeability prevented by HgCl2.

UI MeSH Term Description Entries
D007267 Injections Introduction of substances into the body using a needle and syringe. Injectables,Injectable,Injection
D009865 Oocytes Female germ cells derived from OOGONIA and termed OOCYTES when they enter MEIOSIS. The primary oocytes begin meiosis but are arrested at the diplotene state until OVULATION at PUBERTY to give rise to haploid secondary oocytes or ova (OVUM). Ovocytes,Oocyte,Ovocyte
D011892 Rana catesbeiana A species of the family Ranidae (true frogs). The only anuran properly referred to by the common name "bullfrog", it is the largest native anuran in North America. Bullfrog,Bullfrogs,Rana catesbeianas,catesbeiana, Rana
D002463 Cell Membrane Permeability A quality of cell membranes which permits the passage of solvents and solutes into and out of cells. Permeability, Cell Membrane
D003315 Cornea The transparent anterior portion of the fibrous coat of the eye consisting of five layers: stratified squamous CORNEAL EPITHELIUM; BOWMAN MEMBRANE; CORNEAL STROMA; DESCEMET MEMBRANE; and mesenchymal CORNEAL ENDOTHELIUM. It serves as the first refracting medium of the eye. It is structurally continuous with the SCLERA, avascular, receiving its nourishment by permeation through spaces between the lamellae, and is innervated by the ophthalmic division of the TRIGEMINAL NERVE via the ciliary nerves and those of the surrounding conjunctiva which together form plexuses. (Cline et al., Dictionary of Visual Science, 4th ed) Corneas
D004848 Epithelium The layers of EPITHELIAL CELLS which cover the inner and outer surfaces of the cutaneous, mucus, and serous tissues and glands of the body. Mesothelium,Epithelial Tissue,Mesothelial Tissue,Epithelial Tissues,Mesothelial Tissues,Tissue, Epithelial,Tissue, Mesothelial,Tissues, Epithelial,Tissues, Mesothelial
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D012333 RNA, Messenger RNA sequences that serve as templates for protein synthesis. Bacterial mRNAs are generally primary transcripts in that they do not require post-transcriptional processing. Eukaryotic mRNA is synthesized in the nucleus and must be exported to the cytoplasm for translation. Most eukaryotic mRNAs have a sequence of polyadenylic acid at the 3' end, referred to as the poly(A) tail. The function of this tail is not known for certain, but it may play a role in the export of mature mRNA from the nucleus as well as in helping stabilize some mRNA molecules by retarding their degradation in the cytoplasm. Messenger RNA,Messenger RNA, Polyadenylated,Poly(A) Tail,Poly(A)+ RNA,Poly(A)+ mRNA,RNA, Messenger, Polyadenylated,RNA, Polyadenylated,mRNA,mRNA, Non-Polyadenylated,mRNA, Polyadenylated,Non-Polyadenylated mRNA,Poly(A) RNA,Polyadenylated mRNA,Non Polyadenylated mRNA,Polyadenylated Messenger RNA,Polyadenylated RNA,RNA, Polyadenylated Messenger,mRNA, Non Polyadenylated
D014867 Water A clear, odorless, tasteless liquid that is essential for most animal and plant life and is an excellent solvent for many substances. The chemical formula is hydrogen oxide (H2O). (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed) Hydrogen Oxide
D014982 Xenopus laevis The commonest and widest ranging species of the clawed "frog" (Xenopus) in Africa. This species is used extensively in research. There is now a significant population in California derived from escaped laboratory animals. Platanna,X. laevis,Platannas,X. laevi

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